Systemic therapy of myeloma xenografts by an attenuated measles virus

Abstract

IntroductionAttenuated viruses with tumor specificity have attracted considerable interest as novel anticancer agents, and clinical testing of several such agents is under way. 1 Tumor selectivity of these viruses has been attributed to various intracellular restrictions to their life cycles that are strongly inhibitory to virus propagation in nontransformed cells but that are overridden by cellular factors present in neoplastic cells. [2][3][4][5][6] Measles is an acute viral disease caused by a negative-strand RNA virus of the family Paramyxoviridae, which remains responsible for approximately 1 million deaths each year. 7 In the early stages of measles virus (MV) infection, the lymphoid organs and tissues are predominant sites for viral replication, leading to the formation of giant reticuloendothelial (Warthin-Finkeldey) cells in the tissues. 7 Measles is prevented by the use of a live attenuated virus vaccine now routinely administered during childhood. The Edmonston-B vaccine strain of measles virus (MV-Edm) was attenuated by serial tissue culture passage of a clinical isolate. 7 Despite its profound attenuation as a human pathogen, MV-Edm replicates more efficiently than nonattenuated measles virus in many primate cell lines, inducing cell cell fusion and the formation of characteristic multinucleated syncytia. 8 Here, we compared the ability of MV-Edm to replicate in neoplastic myeloma cells and normal cells, and we investigated its potential as an antitumor agent in human myeloma xenografts in vivo. We report that the virus replicated selectively in a panel of 6 myeloma cell lines and in CD138-sorted myeloma cells from 6 patients and that it caused potent cytopathic effects. When administered intratumorally or intravenously into mice bearing established myeloma xenografts, MV-Edm caused growth inhibition or total regression of 2 different myeloma xenograft models. Materials and methods Cell cultureThe multiple myeloma ARH-77 cell line (ATCC CRL-1621) was obtained from American Type Culture Collection (Rockville, MD). The rest of the multiple myeloma cell lines were kind gifts of Dr John Lust (RPMI 8226), Dr Diane Jelinek (KAS-6/1), and Dr Rafael Fonseca (KMS-11, MM1, JJN-3) from the Mayo Clinic (Rochester, MN). All the human myeloma cell lines were maintained in RPMI-1640 (Gibco BRL, Rockville, MD) supplemented with 10% fetal bovine serum (FBS) except for KAS-6/1, which was grown in media supplemented with 1 ng/mL interleukin (IL)-6 (Sigma, St Louis, MO). Bone marrow aspirates were obtained after institutional review board approval and informed patient consent. Bone marrow samples were drawn into a tube containing heparin and centrifuged on a Ficoll-Hypaque gradient (Amersham Pharmacia, Piscataway, NJ) to enrich for mononuclear cells. Mononuclear cells were then incubated with MACS CD138 microbeads (Miltenyi Biotech, Auburn, CA) for 15 minutes in a 8°C water bath. CD138 ϩ primary myeloma cells were then isolated, washed, and maintained in 10% FBS-RPMI supplemented with 1 ng/mL IL-6. Peripheral blood lym...

Replication of an integrin targeted conditionally replicating adenovirus on primary ovarian cancer spheroids

“…In this paper, we used spheroids for analysis of CRAds, but the model should be applicable to any type of replicating agent. Replication competent viruses currently undergoing preclinical or clinical evaluation for cancer treatment include herpes simplex virus, 65,66 vaccinia, 67 Newcastle disease virus, 68 parvovirus, 69 reovirus, 70 measles virus, 71 retrovirus, 72 influenza virus, 73 poliovirus, 74 vesicular stomatitis virus, 75 and avian sarcoma leucosis virus. 76 Therefore, replication competent viruses represent a dynamic field desperate for practical substrates for primary tumor specimen analyses.…”

Cell carriers to deliver oncolytic viruses to sites of myeloma tumor growth

“…In contrast, primary phytohemagglutinin-stimulated peripheral blood lymphocytes from healthy donors were not efficiently infected with MV-Edm. 24 Severe-combined immunodeficiency mice bearing subcutaneous myeloma xenografts were utilized to address the in vivo application of MV-Edm for the treatment of MM. There was complete tumor regression in severe-combined immunodeficiency mice that were treated with intratumoral injections of MV-Edm but not in tumor bearing mice that were treated with ultraviolet-inactivated MV-Edm.…”

Heat shock protein inhibitors increase the efficacy of measles virotherapy

“…The following primers were applied; RhoA-L63: HIV-1 vectors pseudotyped with VSV.G envelopes were generated by transient transfection of 30 mg transfer vector plasmid, 30 mg pCMV8.91 gag-pol-expressing plasmid and 10 mg pMD.G (envelope vector) into 293T cells plated in T75 flasks using calcium phosphate co-precipitation method and titers were determined as previously described. 3 Assessment of impact of RhoA status on measles-induced fusion MDA-MB231 cells were plated overnight in six-well plates (2 Â 10 5 À10 6 cells per well) and infected with MVCombining measles virus with heat shock protein inhibitors C Liu et al GFP in opti-MEM at MOI 0.5 for 2 h at 37 1C. Media were replaced with fresh growth medium for 2 h at 37 1C, and cells were infected overnight with lentiviruses (MOI 10) in the presence of 8 mg ml À1 polybrene.…”

“…1 Antitumor activity of attenuated strains of measles virus derived from the Edmonston vaccine lineage has been demonstrated in models of lymphoma, 2 multiple myeloma, 3 ovarian cancer, 4 hepatocellular carcinoma, 5 glioblastoma multiforme 6 and breast cancer. 7 After infection of tumor cells, the virus causes a very characteristic cytopathic effect, the development of multinucleated giant cells (syncytia).…”